Homology Between Exon-Containing Portions of Rabbit Genomic Clones for Synovial Cell Collagenase and Human Foreskin and Synovial Cell mRNAs

Fini, M. Elizabeth; Austin, S D; Holt, Peter T.; Ruby, Peggy L.; Gross, Robert; White, Hillary D.; Brinckerhoff, Constance E.

doi:10.1016/s0174-173x(86)80009-7

Cited by 11 publications

(10 citation statements)

References 25 publications

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“…We compared the sequence of rabbit activator with that of rabbit synovial cell collagenase. We found no homology to a 530-bp cDNA clone, designated H9 (6), but this is not surprising since this clone represents primarily the 3'-untranslated portion of the rabbit synovial fibroblast collagenase message (41). Rabbit activator cDNA exhibits weak, but definite, crosshybridization (under reduced stringency) to exoncontaining restriction fragments of a rabbit synovial fibroblast collagenase genomic clone (42).…”

Section: ?He Lys Met -Asp Pro-gly-phe-pro-arg-his-ile-ala-glu-asp-phementioning

confidence: 82%

Cloning of a complementary dna for rabbit proactivator. a metalloproteinase that activates synovial cell collagenase, shares homology with stromelysin and transin, and is coordinately regulated with collagenase

Fini

Karmilowicz

Ruby

et al. 1987

Arthritis & Rheumatism

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Rabbit proactivator is a neutral metalloproteinase that activates another metalloproteinase, procollagenase, and degrades noncollagenous matrix. We describe the construction of an activator complementary DNA (cDNA) clone, which is 1.9 kb, that selects a 2.1-kb messenger RNA (mRNA) in Northern blot hybridizations. Nucleic acid sequence studies of the activator cDNA indicate 1) that it encodes protein M , 53,881, 2) that this protein exhibits -80% homology with rat transin, an oncogene-induced protein with a previously unknown function, and 3) that, in the first 172 residues, it is virtually identical to the rabbit metalloproteinase, stromelysin. Homology between rabbit activator and human skin collagenase is approximately 50%. Activator and collagenase mRNA are coordinately regulated; untreated cultures of rabbit synovial fibroblasts produce low levels of each protein, but addition of phorbol myristate acetate ( W M ) results in an increase in mRNA for both proteins by 2.5-5 hours. Adding alltrans-retinoic acid (lo-%) or dexamethasone (lO-'M) to phorbol-stimulated cells coordinately suppresses both activator and collagenase mRNA. Our data suggest the existence of coordinately regulated metalloproteinases that are important in the modulation of connective tissue metabolism.The metalloproteinase, collagenase (EC 3.2.24.7), is the only neutral proteinase able to initiate degradation of the interstitial collagens, types I, 11, and I11 (for review, see refs. 1 and 2). As such, collagenase plays a crucial role in the degradation of collagen that occurs as a result of both normal physiologic and disease processes (3). To study mechanisms that control the synthesis of collagenase, we have developed a model system of cultured rabbit synovial fibroblasts. These cells are readily available from the synovium of the knees of young rabbits and grow well in culture. Of experimental value is the fact that they make only minimal amounts of collagenase unless stimulated with such agents as interleukin-I , crystals of monosodium urate monohydrate, or the tumor promoter, phorbol myristate acetate (PMA) (r18). Addition of an inducer results in an increase in collagenase messenger RNA (mRNA), which is followed by rapid synthesis and secretion of collagenase protein (6,9). Equally valuable is the fact that collagenase synthesis can be antagonized by glucocorticoids and by retinoids (10,ll). It is believed that both compounds act at the level of transcription to decrease collagenase mRNA and protein, but do not affect the half-life of collagenase mRNA (12). This system has thus allowed us to study both the induction and suppression of collagenase synthesis.Rabbit synovial fibroblast collagenase is synthesized and secreted as an inactive polypeptide, procollagenase ( M , 57,000), with a minor species of glycosylated enzyme (M, 61,000) (9,13). Activation of

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Section: ?He Lys Met -Asp Pro-gly-phe-pro-arg-his-ile-ala-glu-asp-phementioning

confidence: 82%

Cloning of a complementary dna for rabbit proactivator. a metalloproteinase that activates synovial cell collagenase, shares homology with stromelysin and transin, and is coordinately regulated with collagenase

Fini

Karmilowicz

Ruby

et al. 1987

Arthritis & Rheumatism

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“…Expression of MMP1 and MMP3 mRNA in intact rabbit forebrain was studied in Northern blots. MMP‐1 mRNA expression, at the expected size of 2.2 kb [ 5], was strongest in fetal rabbit forebrain, with persistent weak expression in post‐natal immature rabbits, and negligible in adult rabbit forebrain ( Figure 6). An extra band at ≈4 kb likely corresponds to the enterotoxin receptor which is found in the brain endothelium [ 3] and whose mRNA has 19 consecutive bases complementary to the probe.…”

Section: Resultsmentioning

confidence: 99%

Expression of extracellular matrix degrading enzymes during migration of xenografted brain cells

Bigio

Tchélingérian²,

Jacque（摄影）

1999

Neuropathology Appl Neurobio

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Proteolytic enzymes, postulated to create an avenue for cell migration by digestion of host extracellular matrix molecules, have been implicated in neoplastic glial cell migration. A similar process is likely to occur in the developing brain. Fetal rabbit brain fragments transplanted into the striatum of the neonatal Shiverer mouse give rise to cells which migrate from the graft site and differentiate into astrocytes and oligodendrocytes. Proteinase expression by transplanted brain cells was studied using immunohistochemistry and in situ hybridization. Immature donor cells expressed the mRNAs for matrix metalloproteinases (MMP) 1 (collagenase) and 3 (stromelysin). Northern blot analysis of rabbit brain showed that MMP-1 in particular is expressed in the immature rabbit cerebrum and down-regulated during maturation. Immature donor cells exhibited immunoreactivity for urokinase plasminogen activator. However, immunoreactivity was also present in maturing neurons. Donor and host astroglia in the vicinity of grafts were immunoreactive for MMP-2 and tissue-type plasminogen activator. This expression may represent a reactive phenomenon, not specifically related to cell migration, by mature astrocytes. Based upon our findings, MMP-1 appears to be a candidate for involvement in migration of immature brain cells in the cerebrum.

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“…For example, the collagenase cDNA clones do not hybridize to human RNAs that we have tested under conditions of moderate hybridization stringency (Fini ME, Brinckerhoff CE: unpublished data). However, we have already learned that portions of the genomic clones hybridize fairly well to a 2.2 kb transcript from human rheumatoid cells and less well to a 2.2 kb transcript from human foreskin fibroblasts (61).…”

Section: Discussionmentioning

confidence: 99%

Characterization of Rabbit Genes for Synovial Cell Collagenase

Fini

Gross

Brinckerhoff

1986

Arthritis & Rheumatism

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To provide tools for understanding collagenase gene expression in rheumatoid arthritis, we have isolated and characterized genomic clones for rabbit synovial cell collagenase. These clones represent 2 types of collagenase gene, at least 1 of which is transcribed in synovial fibroblasts. By examining the rabbit genome in situ, we provide evidence that there are only 2 different synovial cell collagenase genes found in a haploid genome. Amplification of these genes is not a mechanism far collagenase messenger RNA induction by phorbol esters.The metalloproteinase, collagenase, plays a primary role in the joint destruction seen in rheumatoid arthritis (1). Vast excesses of this enzyme are secreted by the fibroblasts of the rheumatoid pannus. The fact that collagen breakdown can be initiated only by collagenase makes this enzyme rate-limiting in collagenolysis. One approach to the successful control of collagenolysis in diseases such as rheumatoid arthritis may lie in inhibiting the production of collagenase. Thus, an understanding of mechanisms regulating Studies in our laboratory, designed to understand the control of collagenase secretion, have been facilitated by the development of a cell culture system of synovial fibroblasts from rabbits (2). These cultures secrete negligible levels of collagenase when untreated. However, they can be stimulated to secrete major amounts of collagenase by treatment with numerous agents, including the tumor promoter, phorbol myristate acetate (PMA) (3), and crystals of monosodium urate monohydrate (4). Furthermore, additional agents such as retinoic acid or dexamethasone can reverse or suppress induction (5). To date, experiments with human synovial cells have corroborated our findings with the rabbit cell studies, and have thus validated the rabbit system as a model (6,7).Earlier studies demonstrated that inducers act to increase the synthesis of new collagenase protein rather than stimulate the release of previously synthesized enzyme stored intracellularly (8). With the development of a complementary DNA (cDNA) clone for collagenase, we showed that on induction, an increase in immunoreactive collagenase secreted into the culture medium is directly correlated with increased collagenase messenger RNA (mRNA) levels (9). Several steps in RNA metabolism could contribute to the increase in collagenase message levels, including increased transcription from the collagenase gene(s), greater mRNA stability, or changes in processing of pre-mRNA to mRNA. To study the contribution of these processes to mRNA induction, as well as the mechanisms by which they operate, requires more than a cDNA clone, which is merely a copy of mRNA. The collagenase gene itself must be isolated

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Homology Between Exon-Containing Portions of Rabbit Genomic Clones for Synovial Cell Collagenase and Human Foreskin and Synovial Cell mRNAs

Cited by 11 publications

References 25 publications

Cloning of a complementary dna for rabbit proactivator. a metalloproteinase that activates synovial cell collagenase, shares homology with stromelysin and transin, and is coordinately regulated with collagenase

Cloning of a complementary dna for rabbit proactivator. a metalloproteinase that activates synovial cell collagenase, shares homology with stromelysin and transin, and is coordinately regulated with collagenase

Expression of extracellular matrix degrading enzymes during migration of xenografted brain cells

Characterization of Rabbit Genes for Synovial Cell Collagenase

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